RP-HPLC法测定大果山楂果实中四种氨基酸的含量  

Determination of four amino acids in fruit of Malusdoumeri(Bois)Chev

作　　者：祁佳佳 杨嘉怡 范亚梦 李典鹏 王先友[1] QI Jiajia;YANG Jiayi;FAN Yameng;LI Dianpeng;WANG Xianyou(School of Pharmacy,Henan University,Kaifeng 475004,Henan,China;Guangxi Key Laboratory of Functional Phytochemicals Research and Utilization,Guilin 541006,Guangxi,China)

机构地区：[1]河南大学药学院,河南开封475004 [2]广西植物功能物质研究与利用重点实验室,广西桂林541006

出　　处：《化学研究》2022年第6期512-516,共5页Chemical Research

基　　金：广西植物功能物质研究与利用重点实验室开放基金(FPRU2017-3)。

摘　　要：建立了大果山楂中苯丙氨酸、组氨酸、酪氨酸和亮氨酸含量测定的RP-HPLC方法,确定了流动相的组成及比例、提取方式和样品用量等。利用7 mol/L HCl溶液回流提取,减压浓缩、挥干HCl,稀释后测定。采用C_(18)(4.6 mm×250 mm,5μm)色谱柱分离;柱温为30℃;流动相:甲醇与水体积比为3∶97(pH=4);流速1.0 mL/min,进样量为10μL;检测波长:λD=205 nm。苯丙氨酸在0.025~1 g/L范围内,线性关系良好(R^(2)=0.9992),加样回收率为100.1%(RSD=1.7%,n=6);组氨酸在0.025~1 g/L范围内,线性关系良好(R^(2)=0.9988),加样回收率为98.40%(RSD=1.9%,n=6);酪氨酸在0.025~0.8 g/L浓度范围内,线性关系良好(R^(2)=0.9984),加样回收率为97.46%(RSD=1.8%,n=6);亮氨酸在0.1~2.0 g/L范围内具有良好的线性关系(R^(2)=0.9981),加样回收率为96.82%(RSD=1.8%,n=6)。本实验建立的HPLC方法稳定性好、精密度高、重复性和回收率良好,可用于大果山楂中该四种氨基酸含量的简便快速测定。HPLC methods for the determination of phenylalanine,histidine,tyrosine and leucine in fruit of Malusdoumeri(Bois)Chev.were established in this paper.Study contents included the defination of composition and proportion of mobile phase,extraction method and sample usage.Amino acids were extracted by refluxation with 7 mol/L HCl,then concentrated and volatilized under reduced pressure to remove HCl.Separation was performed in C_(18)column(250 mm×4.6 mm,5μm);The column temperature is 30℃;Mixture of methanol and water 3∶97(pH=4)was used as mobile phase with flow rate at 1.0 mL/min.Injection volume was 10μL.The detection wavelengths were set at 205 nm.The linear relationship of phenylalanine was good(R^(2)=0.9992)at range of 0.025-1 g/L.Average recovery of phenylalanine was 100.1%,with RSD=1.7%(n=6).The linear relationship of histidine was good(R^(2)=0.9988)at range of 0.025-1 g/L.The average recovery of histidine was 98.43%,with RSD=1.9%(n=6).The linear relationship of tyrosine was good(R^(2)=0.9984)in the range of 0.025-0.8 g/L,and the recovery was 97.46%(RSD=1.8%,n=6).Leucine had a good linear relationship(R^(2)=0.9981)at the range of 0.1-2.0 g/L,and the recovery was 96.82%(RSD=1.8%,n=6).The HPLC method established in this experiment has good stability,high precision,good repeatability and recovery.It can be used for the determination of these four amino acids in fruit of Malusdoumeri(Bois)Chev simply and rapidly.

关 键 词：大果山楂 氨基酸 反相高效液相色谱法 含量测定 

分 类 号：O657[理学—分析化学]